Steroid sulfatase inhibitors and methods for making and using the same

ABSTRACT

Novel sulfatase inhibitor compounds useful in the treatment of estrogen dependent illnesses are disclosed. These compounds generally comprise a steroid nucleus substituted at the C17 position. Methods for synthesizing these compounds and using them in the therapeutic and/or prophylactic treatment of a patient are also disclosed.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a divisional of U.S. Ser. No. 09/490,302, filed Jan.24, 2000, which is a continuation-in-part of U.S. Ser. No. 09/236,842,filed Jan. 25, 1999, now U.S. Pat. No. 6,288,050, which is acontinuation-in-part of U.S. Ser. No. 08/897,247, filed Jul. 18, 1997,now U.S. Pat. No. 5,880,115.

FIELD OF THE INVENTION

The present invention relates to sulfatase inhibitors and methods formaking and using the same. These methods include use of these compoundsin therapeutic and prophylactic treatments for estrogen dependentillnesses.

BACKGROUND OF THE INVENTION

Estrogen levels in breast tumors of post-menopausal women are at leastten times higher than estrogen levels in plasma. The high levels ofestrogen in these tumors are due to in situ formation of estrogen,possibly through conversion of is estrone sulfate to estrone by theenzyme estrone sulfatase. Inhibitors of estrone sulfatase are thereforepotential agents for the treatment of estrogen-dependent breast cancers.Most estrone sulfatase inhibitors are steroidal in nature. Althoughestrone-3-O-sulfamate (EMATE) is believed to be the most potentinhibitor of estrone sulfatase, recent evidence indicates that thiscompound is a potent estrogen. This compound is therefore not useful inthe treatment of estrogen dependent illnesses.

Reed and co-workers reported on sulfatase inhibitory activities ofestrone-3-O-methylthiophosphonate, estrone-3-O alkyl and arylsulfonates, estrone-3-O-phosphonates and thiophosphonates and estronesulfamates in: Duncan et al., “Inhibition of Estrone Sulfatase Activityby Estrone-3-methylthiophosphonate”, Cancer Res. 53:298-303 (1993);Howarth et al., “Phosphonates and Thiophosphonates as SulfateSurrogates: Synthesis of Estrone-3-methylthiophosphonate, a PotentInhibitor of Estrone Sulfatase”, Bioorg. Med. Chem. Lett. 3:313-318(1993); Howarth et al., “Estrone Sulfamates: Potent Inhibitors ofEstrone Sulfatase with Therapeutic Potential”, J. Med. Chem. 37:219-221(1994); and Purohit et al., “In vivo Inhibition of Oestrone Sulphataseand Dehydroepiandrosterone Sulphatase by Oestrone-3-O-sulphamate”, Int.J. Cancer, 63:106-111 (1995).

Li and co-workers reported the synthesis and sulfatase inhibitoryactivities of sulfonate and its analogues, methylene sulfonates andphosphate that contain the estrone nucleus in Li et al., “Synthesis andBiochemical Studies of Estrone Sulfatase Inhibitors”, Steroids,58:106-111 (1993); Dibbelt et al., “Inhibition of Human PlacentalSterylsulfatase by Synthetic Analogs of Estrone Sulfate”, J. SteroidBiochem. Molec. Biol., 50(5/6):261-266 (1994); and Li et al., “EstroneSulfate Analogs as Estrone Sulfatase Inhibitors”, Steroids60:299-306(1995). Estrone-3-amino derivatives are reported in Selcer et al.,“Inhibition of Placental Estrone Sulfatase Activity and MCF-7 BreastCancer Cell Proliferation by Estrone-3-amino Derivatives”, J. SteroidBiochem. Molec. Biol., 59(1):83-91 (1996).

U.S. Pat. No. 5,567,831 is directed to the use of non-steroidalsulfatase inhibitor compounds in the treatment of estrogen dependentillnesses.

U.S. Pat. No. 5,571,933 is directed to derivatives of estra1,3,5(10)trien-17-one, 3-amino compounds and methods for using thesecompounds in the treatment of estrogen dependent illnesses.

U.S. Pat. Nos. 5,556,847 and 5,763,492, are directed to steroidal andnon-steroidal sulfatase inhibitors, respectively, and methods for usingthese inhibitors to effect memory enhancement. Use of these inhibitorsin the treatment of estrogen dependent illnesses is not disclosed.

U.S. Pat. Nos. 5,616,574 and 5,604,215 disclose steroid sulphataseinhibitors and the methods of using the same. The disclosed compoundsare potent estrogens and metabolize to form estrones, in contrast to thecompounds of the present invention.

U.S. Pat. No. 5,763,432 discloses steroid inhibitors of estronesulfatase; the patent does not appear to disclose the compounds of thepresent invention, or compounds in which the substituent at the C17position of a steroid nucleus interacts with a lipid bilayer.

There remains a need for potent sulfatase inhibitors that aremetabolically stable, more selective, and devoid of estrogenic activity.

SUMMARY OF THE INVENTION

The present invention has met the above described need by providingnon-estrogenic compounds useful as steroid sulfatase inhibitors. Thesecompounds generally comprise a substituted steroid ring system having 4attached rings; in the compounds of the present invention this structureis generally depicted by formula 1:

wherein X and Y are both carbons and the bond between X and Y is eithersingle or double, as is further described below. The compounds of thepresent invention can is be generally described as sulfatase inhibitors,and derive this inhibition ability from the presence of an

group. The nitrogen in this group can be further substituted with one ormore hydrogen atoms, one or more lower alkyl groups having 1 to 6carbons, or combinations thereof. Thus, the present compounds comprise asulfamate group or a 6-membered cyclic sulfamate group attached to the“A” ring of the steroid nucleus, thereby resulting in a compound with 5attached rings. More specifically, the group can be attached to the “A”ring in the steroid nucleus in which case a sulfamate ester of thesteroid would be represented. Alternatively, the “N”, “S”, and the “O”attached to the S by a single bond can, together with a carbon attachedto the N, form a fifth ring adjacent to the “A” ring of the steroidnucleus. The bond between the N and the carbon could be double, in whichcase an oxathiazine dioxide would be represented, or single, in whichcase a dihydro-oxathiazine dioxide ring would be represented.

The present invention is also directed to methods for synthesizing thesteroid sulfatase inhibitors disclosed herein.

In addition, the present invention relates to methods for using thesecompounds as sulfatase inhibitors. These methods generally compriseincorporating one or more of the compounds into a suitablepharmaceutical carrier and administering a therapeutically orprophylactically effective amount of the compound to a patient.

It is an aspect of this invention to provide compounds for inhibitingthe steroid sulfatase enzyme produced in the body.

It is a further aspect of the invention to provide estrone sulfataseinhibitor compounds having anti-tumor or synergistic activity withanti-estrogen and aromatase inhibitors.

It is a further aspect of the present invention to provide estronesulfatase inhibitor compounds having activity against estrogen dependentillnesses.

Yet another aspect of the invention is to provide methods fortherapeutically or prophylactically treating a patient with thesulfatase inhibitor compounds of the present invention.

It is another aspect of this invention to provide derivatives ofsulfatase inhibitor compounds that are not metabolized to compounds thatare estrogenic.

These and other aspects of the invention will be more fully understoodto those skilled in the art upon review of the following description andappended claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 generally illustrates the estrone sulfatase pathway.

FIG. 2 illustrates the manner in which a C17 substituent, represented by“R₃” in the Figure, interacts with a lipid bilayer.

FIG. 3 illustrates a scheme for preparing a compound of Class 1,according to the methods of Example 1.

FIG. 4 illustrates a scheme for preparing compounds of Class 2,according to the methods of Example 2.

FIG. 5 illustrates a scheme for preparing compounds of Class 2,according to the methods of Example 2.

FIG. 6 illustrates a scheme for preparing compounds of Class 2,according to the methods of Example 2.

FIG. 7 illustrates a scheme for preparing a compound of Class 3,according to the methods of Example 3.

FIG. 8 illustrates a scheme for preparing a compound of Class 4.

FIG. 9 illustrates a reaction scheme for preparing a compound of Class10, according to the methods of Example 5.

FIG. 10 illustrates a reaction scheme for preparing a compound of Class12.

FIG. 11 illustrates a scheme for preparing compounds of Class 2,according to the methods of Example 4.

FIG. 12 illustrates a scheme for preparing a compound of Class 6,according to the methods of Example 6.

DETAILED DESCRIPTION OF THE INVENTION

As used herein, the term “patient” refers to members of the animalkingdom including but not limited to human beings.

The present invention is directed to compounds having formula 2:

wherein R₁ and R₂ are independently selected from hydrogen and a loweralkyl group having one to six carbons;

R₃ is selected from the group:

 R₄ and R₅ are independently selected from hydrogen, straight orbranched chain alkyl groups having one to fourteen carbons, and straightor branched chain alkoxy groups having one to six carbons;

R₆ is selected from hydrogen and straight or branched chain alkyl groupshaving one to fourteen carbons;

R₇ is selected from

hydrogen and straight or branched chain alkyl groups having one tofourteen carbons, R₁₂ and R₁₃ are independently selected from hydrogen,straight or branched chain alkyl groups having one to fourteen carbonsand straight or branched chain alkoxy groups having one to six carbons,and R₁₄ is selected from hydrogen, and straight or branched chain alkylgroups having one to fourteen carbons;

R₈ is selected from hydrogen and straight or branched chain alkyl groupshaving one to six carbons;

R₉ is selected from straight or branched chain alkanoyl groups havingone to fifteen carbons, straight or branched chain alkyl groups havingone to fourteen carbons, CO(CH₂)_(m)CH₃ and COR₁₀;

R₁₀ is a straight or branched chain alkyl group having one to fourteencarbons, but the alkyl group is not a branched chain when the alkylgroup is a C₁ or C₂ alkyl group; m is 0 to 2;

R₁₁ is a branched chain alkyl group having three to fourteen carbons;and

X and Y are both carbons and the bond between X and Y is either singleor double, except when R₃ is

the bond between X and Y is single.

As will be appreciated by those skilled in the art, these compounds areestrones, more particularly 1,3,5(10) trienes. Suitable steroid ringsystems include the substituted estrones:

2-OH-estrone

2-methoxy-estrone

4-OH-estrone

6 alpha-OH-estrone

7 alpha-OH-estrone

7 alpha-alkylarnido-estrone

16 alpha-OH-estrone

16 beta-OH-estrone

The present invention is further directed to compounds having theformula (3):

wherein R₃ is selected from

 R₄ and R₅ are independently selected from hydrogen, straight orbranched chain alkyl groups having one to fourteen carbons, and straightor branched chain alkoxy groups having one to six carbons;

R₆ is selected from hydrogen and straight or branched chain alkyl groupshaving one to fourteen carbons;

R₇ is selected from

hydrogen and straight or branched chain alkyl groups having one tofourteen carbons, R₁₂ and R₁₃ are independently selected from hydrogen,straight or branched chain alkyl groups having one to fourteen carbonsand straight or branched chain alkoxy groups having one to six carbons,and R₁₄ is selected from hydrogen, and straight or branched chain alkylgroups having one to fourteen carbons;

R₈ is selected from hydrogen and straight or branched chain alkyl groupshaving one to six carbons;

R₉ is selected from straight or branched chain alkanoyl groups havingone to fifteen carbons, straight or branched chain alkyl groups havingone to four teen carbons, CO(CH₂)_(m)CH₃ and COR₁₀;

R₁₀ is a straight or branched chain alkyl group having one to fourteencarbons, but the alkyl group is not a branched chain when the alkylgroup is a C₁ or C₂ alkyl group; m is 0 to 2;

R₁₁ is a branched chain alkyl group having three to fourteen carbons; Xand Y are both carbons and the bond between X and Y is either single ordouble, except when R₃ is

the bond between X and Y is single; and

K is nitrogen and L is carbon and the bond between K and L is eithersingle or double.

For the compounds represented by Formulas 2 and 3, “X”, “Y” and “L” areall carbon, and “K” is nitrogen; the bond connecting X and Y, and K andL can generally be either a single bond or a double bond. It will beappreciated that when the bond is single, the X and K have a hydrogenattached thereto, and the Y and L have two hydrogens attached thereto.

Numerous estrones are therefore the subject of the present invention.For ease of reference, these compounds, which generally have eitherformula 2 or formula 3 as described above, will be referred to herein asClass 1 through Class 12 compounds depending on the “R₃” attachment, asidentified in Table 1 below: Class 1 through Class 6 refer to compoundsof formula 2 and Class 7 through Class 12 refer to compounds of formula3.

TABLE 1 Class for Formula Class for 2 Formula 3 R₃ Class 1 Class 7

Class 2 Class 8

Class 3 Class 9

Class 4 Class 10

Class 5 Class 11

Class 6 Class 12

Preferably, R₁ and R₂ are both hydrogen. Further preferred embodimentsinclude compounds of Classes 1 and 7 wherein m is equal to 2; compoundsof Classes 2 and 8 wherein R₄ and R₅ are both CH₃; compounds of Classes2 and 8 wherein R₄ is hydrogen and R₅ is a straight or branched chainalkyl group having one to eight carbons or a straight or branched chainalkoxy group having one to three carbons; compounds of Classes 2 and 8wherein R₄ is a straight or branched chain alkyl group having one tofive carbons and R₅ is a straight or branched chain alkyl group havingone to six carbons; compounds of Classes 3 and 9 wherein R₆ is astraight or branched chain alkyl group having one to eight carbons,especially CH₂CH₃; compounds of Classes 4 and 10 wherein R₇ is astraight or branched chain alkyl group having one to eight carbons,especially a straight chain alkyl group having three carbons (propyl);compounds of Classes 4 and 10 wherein R₇ is

R₁₂ and R₁₃ are both hydrogen, methyl, propyl or isopropyl; or R₁₂ isethyl and R₁₃ is isopropyl; compounds of Classes 5 and 11 wherein R₁₁,is a straight or branched chain alkyl group having three to eightcarbons, especially a branched chain alkyl group having three carbons(isopropyl); compounds of Classes 6 and 12 wherein R₈ is hydrogen or astraight or branched chain alkyl group having one to six carbons and R₉is a straight or branched chain alkyl group having two to eight carbonsor a straight or branched chain alkanoyl group having one to eightcarbons; and compounds of Classes 6 and 12 wherein R₈ is hydrogen and R₉is a a straight or branched chain alkanoyl having four carbons.

The compounds of the present invention are useful as sulfataseinhibitors. These compounds generally include a substituted steroidnucleus The substituent is attached to the steroid nucleus at the C17position of the nucleus. This is represented in Formulas 2 and 3 as the“R₃” group. Because of the unique substituents found at the C17 positionof the steroid nucleus in the present compounds, the steroidal moleculereleased after inactivation of the enzyme is not estrogenic. Thecompounds of the present invention function as active-site directedirreversible inhibitors of estrone sulfatase. The sulfamate grouprecognizes and binds to the steroid sulfatase or estrone sulfatase inthe estrone sulfatase pathway, thereby preventing the conversion ofestrone sulfate to estrone. The estrone sulfatase pathway is shown inFIG. 1.

In addition, the unique substituents at the C17 position of the steroidnucleus provide enhanced sulfatase inhibition when compared to compoundslacking these substituents. Since estrone sulfatase is a membrane boundenzyme, the substituents on the C17 position as described herein providefor a hydrophobic interaction between the substituent and the lipidbilayers of various membranes of the patient. This interaction resultsin an increase in the binding affinity of the inhibitor to the enzyme.As illustrated in FIG. 2, the C17 substituent (R₃) interacts with thetwo layers (14 and 16) of the lipid bilayer of a membrane. Hydrophobicinteraction keeps the substituent within the bilayer. This additionalbinding site results in more potent sulfatase inhibitory activity.

The present invention is further directed to the synthesis of theabove-described compounds. Various reaction schemes are illustrated inFIGS. 3-12.

FIG. 3 outlines the synthesis of a compound of formula 2 when R₁ and R₂are both equal to hydrogen and R₃ equals

with m equal to 2. This is a compound of Class 1. FIGS. 4, 5 and 6illustrate the synthesis of compounds of Class 2. The particularcompounds synthesized in FIG. 4 are generally represented by formula 2wherein R₁ and R₂ are both hydrogen, R₃ is

and R₄ is hydrogen, methyl, ethyl, propyl or isopropyl and R₅ is any ofthe “R₅” groups corresponding with compounds 11a-11s in FIGS. 4, 5 and6.

FIG. 7 shows a method of synthesizing a compound of Class 3, which isgenerally represented by formula 2 above, wherein R₃ is

The particular embodiment of the Class 3 compound shown in FIG. 7 isrepresented by formula 2 wherein R₁ and R₂ are both hydrogen and R₆ is astraight chain alkyl group having 3 carbons. FIG. 8 depicts thesynthesis of a compound of Class 4, generally represented by formula 2above wherein R₃ equals

The particular embodiment of the Class 4 compound shown in FIG. 8represents formula 2 wherein R₁ and R₂ are both hydrogen and R₇ is astraight chain alkyl group having 3 carbons. It will be understood bythose skilled in the art that the Figures depict specific embodiments ofthe various compounds generally represented by formula 2; othercompounds within formula 2 having different “R” groups or different “m”values can easily be prepared by those skilled in the art by followingthe schemes generally outlined in the Figures and making the necessarysubstitutions.

FIGS. 9 and 10 show synthesis schemes for preparation of compounds ofClass 10 and Class 12. More specifically, FIG. 9 represents thesynthesis of a Class 10 compound, which is represented by formula 3wherein R₃ is equal to

R₁₂ is hydrogen, and R₁₃ is a straight chain alkyl group having 3carbons (propyl). FIG. 10 represents the synthesis of Class 12compounds, represented by formula 3 above wherein R₃ is

R₈ is hydrogen and R₉ is a straight chain alkanoyl group having 3carbons. Again, by following the general synthesis schemes depicted inFIGS. 9 and 10, compounds generally represented by formula 3 abovehaving a variety of substituents can be prepared by using the necessarycompounds to obtain the desired sulfatase inhibitor.

FIG. 11 shows a synthesis scheme for preparation of Class 2 compounds.The particular compounds synthesized in FIG. 11 are generallyrepresented by formula 2 wherein R₁ and R₂ are both hydrogen, R₃ is

R₄ is hydrogen or methyl and R₅ is one of the “R₅” groups correspondingwith compounds 29d, 29e, 29f, 29g, 29h and 29m in FIG. 11.

FIG. 12 shows a synthesis scheme for preparation of Class 6 compounds.The particular compound synthesized in FIG. 12 is generally representedby formula 2 wherein R₁ and R₂ are both hydrogen, R₃ is

R₈ is hydrogen and R₉ is a butyryl group.

The present invention is further directed to methods for using thecompounds described above to therapeutically and/or prophylacticallytreat a patient for an estrogen dependent illness. Such illnessesinclude, but are not limited to, breast cancer, vaginal cancer,endometrial cancer, ovarian cancer and endometriosis.

The methods of the present invention include the steps of: a)incorporating one or more of the compounds of the present invention in asuitable pharmaceutical carrier; and b) administering either atherapeutically effective dosage or a prophylactically effective dosageof the compounds incorporated in the carrier to a patient.

The term “suitable pharmaceutical carrier” refers to any carrier knownin the pharmaceutical arts for administration of compounds to a patient.Any suitable pharmaceutical carrier can be used, so long ascompatibility problems do not arise. A preferred pharmaceutical carrieris physiologic saline (0.9% sodium chloride), 95% dextrose in water.

Administration of an effective dosage or effective amount of thesulfatase inhibitor to a patient can be accomplished by parenteralinjection, such as intravenously, intrathecally, intramuscularly orintra-arterially. The compounds can also be administered orally ortransdermally, or by any other means known to those skilled in the art.Oral administration is preferred.

As used herein, the terms “effective amount” and “effective dosage”refer to that amount of one or more of the compounds disclosed hereinrequired to achieve at least some level of enzyme inhibition in thepatient. Typically, this will be an amount sufficient to impart adesired result or a desired treatment in a patient, such as reducingtumor size or lowering estrogen concentration. The term “therapeuticallyeffective amount” refers to that amount of one or more of the compoundsof the present invention required to therapeutically treat a patient.Such treatment is appropriate in patients having an estrogen-dependentillness. Similarly, the term “prophylactically effective amount” refersto that amount of one or more of the compounds of the present inventionneeded to prophylactically treat a patient. Such treatment isappropriate in patients who, for example, undergo surgery to removecancerous growths; the compounds of the present invention would beadministered to inhibit growth of any new tumor cells which appear. Itwill be appreciated that there is overlap between “therapeutic” and“prophylactic” treatment.

As will be appreciated by those skilled in the art, the dosage ofcompound given, the route of administration and the duration of therapywill be dependent upon the individual being treated, taking intoconsideration such factors as the particular estrogen dependent illnessbeing treated, the body weight of the patient, other therapies beingemployed to treat the patient, and the condition, clinical response andtolerance of the patient. Dosage, administration, and duration oftherapy can be determined by one skilled in the art upon evaluation ofthese and similar factors. A typical patient will be a post-menopausalfemale or pre-menopausal female who has been ovariectomized. Althoughthe dosage and administration will vary from patient to patient, atypical dose will range between 0.005 mg and 2 mg of the presentcompounds per kg of body weight, and will be administered daily.

EXAMPLES

The following examples are intended to illustrate the invention andshould not be construed as limiting the invention in any way.

For all of the examples, chemicals and silica gel were purchased fromAldrich Chemical Company (Milwaukee, Wis.). The chemicals were checkedfor purity by thin layer chromatography and NMR. Biochemicals, estroneand estrone sulfate were obtained from Sigma Chemical Company (St.Louis, Mo.). [6,7-³3H]Estrone sulfate was purchased from Dupont Company.Melting points were determined on a Thomas Hoover capillary meltingpoint apparatus and were uncorrected. Proton NMR spectra were obtainedwith a Bruker WH-300 (300 MHz) spectrophotometer. Elemental analyseswere performed by Atlantic Microlab Inc. (Norcross, Ga.). Radioactivesamples were analyzed with Packard Tri-Carb 4530 and Beckman LS-6500Liquid Scintillation Counters. The liquid scintillation cocktail wasEcolume (ICN, Costa Mesa, Calif.), and Packard Utima Gold.

Example 1 Preparation of Class 1 Compounds

Reference numerals correspond with those shown in FIG. 3.

Synthesis of 3-Benzyloxyestrone (Compound 2)

To a solution of estrone (compound 1) comprising 5 g (22 mmol) ofcompound 1 in acetone (150 ml) was added K₂CO₃ (4.56 g, 33 mmol) andbenzyl bromide (3.9 ml, 33 mmol). The solution was refluxed for 2 daysand the solution was evaporated to dryness and extracted with ethylacetate. The ethyl acetate layer was dried (Na₂SO₄) and the solution wasevaporated to dryness to give a yellowish solid. Excess benzyl bromidein the solid was washed out with petroleum ether to give a white solid(compound 2). The solid was used for the next step without furtherpurification (˜81% yield).

Synthesis of3-Benzyloxy-17-(trinluoromethanesulfonyl)estra-1,3,5(10),16-tetraene(Compound 3)

To a solution of compound 2, comprising 4.8 g (13 mmol) of compound 2 inCH₂Cl₂ (70 ml) was added 2,6-di-tert-butyl-4-methylpyridine (3 g, 14mmol) and triflic anhydride (6 g, 27 mmol) at 0° C. The mixture wasstirred for 5 hrs at room temperature and the mixture was filtered andthe filtrate was washed with 10% sodium bicarbonate solution. Thesolution was then dried (Na₂SO₄), filtered and evaporated andchromatographed (petroleum ether:ethyl acetate, 2:1) to give a yellowishsolid (5.2 g, 84.5%) (compound 3).

Synthesis of3-Benzyloxy-17-(N-propylcarbamoyl)estra-1,3,5(10),16-tetraene (Compound4)

A mixture of triflate compound 3 (1.4 g, 3.02 mmol), palladium (II)acetate (60 mg, 0.26 mmol), 1,3-Bis(diphenylphosphino)propane (dppp)(110 mg, 0.26 mmol), triethylamine (2 ml) and n-propylamine (7 ml) indimethylformamide (DMF, 20 ml) was heated at 70° C. with carbon monoxidebubbled through for 5.5 hours. The reaction mixture was then dilutedwith ethyl acetate and washed with 10% aqueous hydrochloric acid (HCl),10% aqueous sodium bicarbonate (Na₂HCO₃) and brine. The organic layerwas dried with sodium sulfate (Na₂SO₄), concentrated and the residue waspurified by silica gel chromatography and eluted with petroleumether:ethyl acetate (EtOAc), (4:1), yielding the pure α,β-unsaturatedamide (compound 4) (1.04 g, 80%). m.p. 136-138° C.; ¹H NMR (300 MHz,CDCl₃) δ 0.92 (t, 3H, CH₂CH₃), 0.99 (s, 3H, CH₃), 3.22-3.29 (q, 2H,NHCH₂), 5.0 (s, 2H, CH₂Ph), 5.63 (brs, 1H, NH), 6.30 (brs, 1H, 16-CH),6.70 (d, 1H, ArH), 6.74-6.78 (dd, 1H, ArH), 7.17 (d, 1H, ArH), 7.29-7.42(m, 5H, CH₂Ph).

Synthesis of 3-Hydroxy-17-(N-propylcarbamoyl)estra-1,3,5(10),16-tetraene(Compound 5)

Compound 4 (0.9 g, 2.09 mmol) was dissolved in chloroform (30 ml) andtrimethylsilyl iodide (TMS-I, 0.9 ml, 6.28 mmol) was added in oneportion to the reaction mixture at room temperature and the solution wasstirred at room temperature for 3 hrs. Methanol (15 ml) was added to thesolution to quench the reaction and the methanol was evaporated underreduced pressure. The remaining chloroform solution was washed with 10%sodium thiosulfate solution to neutralize the iodide and the solutionwas dried with sodium sulfate (Na₂SO₄). The solution was evaporated andchromatographed using gradient elution from plain CH₂Cl₂ to 10% ethylacetate in CH₂Cl₂ yielding 605 mg of compound 5 (85.1 % yield). m.p.168.5-169° C.; ¹HNMR (300 MHz, CDCl₃) δ 0.92 (t, 3H, CH₂CH₃), 0.99 (s,3H, CH₃), 3.23-3.29 (q, 2H, NHCH₂), 4.66 (s, 1H, OH), 5.64 (brs, 1H,NH), 6.30 (brs, 1H, 16-CH), 6.55 (d, 1H, ArH), 6.59-6.63 (dd, 1H, ArH),7.12 (d, 1H, ArH).

Synthesis of Compound 6

To a solution comprising 400 mg (0.93 mmol) of compound 5 and2,6-di-tert-butyl4-methylpyridine (DBMP) (0.6 g, 2.79 mmol) in CH₂Cl₂(30 ml) was added sulfamoyl chloride (1 g, 8.6 mmol) portionwise withstirring at 0° C. The solution was stirred for 3.5 hrs at roomtemperature. The solution was washed with water until neutral, driedover Na₂SO₄, and evaporated under reduced pressure. The solid obtainedwas purified by chromatography (CH₂Cl₂:petroleum ether:ethyl acetate,2:2:1) to give 404 mg of compound 6 (82.7%). m.p. 185.3-186° C.; ¹H NMR(300 MHz, DMSO-d₆) δ 0.83 (t, 3H, CH₂CH₃), 0.91 (s, 3H, CH₃), 3.04 (t,2H, NHCH₂), 6.34 (brs, 1H, 16-CH), 6.98 (d, 1H, ArH), 6.99-7.02 (dd, 1H,ArH), 7.32 (d, 1H, ArH), 7.74 (brs, 1H, NHCH₂), 7.89 (s, 2H, NH₂).Analysis calculated for C₂₂H₃₀N₂O₄S: C, 63.13; H, 7.22; N, 6.69. FoundC, 63.36; H, 7.24; N, 6.63.

Synthesis of 17β-(N-Propylcarbamoyl)estra-1,3,5(10)-trien-3-yl sulfamate(7)

A solution comprising 108 mg of compound 6 in 7 ml of ethanol wasstirred over 15 mg of palladium (10 wt. % on activated carbon, 50% wetbasis) under an atmosphere of hydrogen for 5h. The catalyst was removedby filtration with diatomaceous earth and the filtrate was concentrated.The residue was purified by chromatography on silica gel (hexane:EtOAc,1:1), yielding pure compound 7 (87 mg, 80%). ₁H NMR (270 MHz, DMSO-d₆) δ0.59 (s, 3H, CH₃), 0.84 (t, 3H, J=7.3 Hz, CH₂CH₃), 2.90(dt, 2H, J=5.8,7.3 Hz, NHCH₂), 3.07-3.20 (m, 1H, 17-CH), 6.95 (d, 1H, J=2.6 Hz, ArH),7.00 (dd, 1H, J=2.6, 8.6 Hz, ArH), 7.33 (d, 1H, J=8.6 Hz, ArH), 7.46(brt, 1H, J=5.8 Hz, NHCH₂), 7.83 (brs, 2H, NH₂).

Example 2 Preparation of Class 2 Compounds

Reference numerals correspond with those shown in FIGS. 4, 5 and 6.

Synthesis of 17-N-butylcarbamoyl)-3-(methoxy)estra-1,3,5(10),16-tetraene(9a)

Oxalyl chloride 0.84 ml was added to a solution comprising 750 mg ofcompound 8 (described in Tetrahedron Letters, 26, 1109-1112, 1985) in 25ml of anhydrous CH₂Cl₂ at 0° C. The solution was stirred at roomtemperature for 5 hours. After removal of the solvent and oxalylchloride, 20 ml of anhydrous tetrahydrofuran (THF) was added to thereaction mixture. The solution of 4 ml was added to a solutioncomprising 0.19 ml of butylamnine in 2 ml of THF at 0° C. and thesolution was stirred for 2 hours. After evaporation of the solvent, 5 mlof water was added. The precipitate formed was filtered and trituratedwith 3 ml of isopropyl ether, yielding compound 9a (124 mg, 70%).

Synthesis of17-(N-Butylcarbamoyl)-3-(hydroxy)estra-1,3,5(10),16-tetraene (10a)

To a solution comprising 119 mg of compound 9a in 2 ml of anhydrousCH₂Cl₂ was added 0.64 ml of BBr₃ (1M solution in CH₂Cl₂) at −15° C.under nitrogen atmosphere. The reaction solution was stirred at roomtemperature for 1.3 hours and quenched by adding methanol and water at0° C. The organic layer was separated and the aqueous layer wasextracted with chloroform (CHCl₃). The combined organic layer was driedwith Na₂SO₄ and concentrated in vacuo. The residue was triturated with2.5 ml of EtOAc and 2.5 ml of hexane, yielding compound 10a (89 mg,78%).

Synthesis of 17-(N-Butylcarbamoyl)estra-1,3,5(10),16-tetraen-3-ylsulfamate (11a)

Sodium hydride (20 mg) was added to a solution comprising 73 mg ofcompound 10a in 2 ml of anhydrous DMF at 0° C. The solution was stirredfor 25 minutes and 126 mg of chlorosulfonamide was added in one portion.The solution was then stirred at room temperature for 2 hours. Ice andsaturated sodium bicarbonate solution was added to the reaction mixture.The precipitate formed was filtered and washed with water. The residuewas purified by chromatography on silica gel (CHCl₃:methanol, 50:1),yielding compound 11a (85 mg, 96%). FAB-MS mn/z 433 (M+H)⁺; ¹H NMR (270MHz, DMSO-d₆) δ 0.88 (t, 3H, J=7.5 Hz, CH₂CH₃), 0.91 (s, 3H, CH₃),3.08(dt, 2H, J=5.6, 6.6 Hz, NHCH₂), 6.34 (s, 1H, 16-CH), 6.97 (s, 1H,ArH), 7.01 (dd, 1H, J=2.6, 8.6 Hz, ArH), 7.33 (d, 1H, J=8.6 Hz, ArH),7.71 (t, 1H, J=5.6 Hz, NHCH₂), 7.88 (s, 2H, NH₂).

Synthesis of 17-(N-Pentylcarbamoyl)estra-1,3,5(10),16-tetraen-3-ylsulfamate (11b)

In similar manners to those described for the synthesis of sulfamate11a, compound 11b was prepared from compound 8. FAB-MS m/z 447 (M+H)⁺;¹H NMR (270 MHz, DMSO-d₆) δ 0.87 (t, 3H, J=6.7 Hz, CH₂CH₃), 0.91 (s, 3H,CH₃), 3.07 (dt, 2H, J=5.8, 6.6 Hz, NHCH₂), 6.34 (s, 1H, 16-CH), 6.97 (s,1H, ArH), 7.01 (dd, 1H, J=2.3, 8.6 Hz, ArH), 7.33 (d, 1H, J=8.6 Hz,ArH), 7.71 (t, 1H, J=5.8 Hz, NHCH₂), 7.88 (s, 2H, NH₂).

Synthesis of 17-(N-Hexylcarbamoyl)estra-1,3,5(10),16-tetraen-3-ylsulfamate (11c)

In similar manners to those described for the synthesis of sulfamate11a, compound 11c was prepared from compound 8. FAB-MS m/z 461 (M+H)⁺;¹H NMR (270 MHz, DMSO-d₆) δ 0.86 (t, 3H, CH₂CH₃), 0.91 (s, 3H, CH₃),3.07 (dt, 2H, NHCH₂), 6.34 (s, 1H, 16-CH), 6.97 (s, 1H, ArH), 7.01 (d,1H, J=8.6 Hz, ArH), 7.33 (d, 1H, J=8.6 Hz, ArH), 7.71 (brt, 1H, NHCH₂),7.88 (s, 2H, NH₂).

Synthesis of 17-(N-Isobutylcarbamoyl)estra-1,3,5(10),16-tetraen-3-ylsulfamate (11d)

In similar manners to those described for the synthesis of sulfamate11a, compound 11d was prepared from compound 8. FAB-MS m/z 433 (M+H)⁺;¹H NMR (270 MHz, DMSO-d₆) δ 0.85 (d, 6H, J=6.9 Hz, CH(CH₃)₂), 0.92 (s,3H, CH₃), 1.73(m, 1H, CH(CH₃)₂), 2.78-3.00 (m, 2H, NHCH₂), 6.36 (s, 1H,16-CH), 6.97 (d, 1H, J=2.3 Hz, ArH), 7.01 (dd, 1H, J=2.3, 8.6 Hz, ArH),7.33 (d, 1H, J=8.6 Hz, ArH), 7.74 (t, 1H, J=6.3 Hz, NHCH₂), 7.88 (s, 2H,NH₂).

Synthesis of 17-(N-tert-Butylcarbamoyl)estra-1,3,5(10),16-tetraen-3-ylsulfamate (11e)

In similar manners to those described for the synthesis of sulfamate11a, compound 11e was prepared from compound 8. FAB-MS m/z 433 (M+H)⁺;¹H NMR (270 MHz, DMSO-d₆) δ 0.92 (s, 3H, CH₃), 1.29 (s, 9H, C(CH₃)₃),6.30 (s, 1H, 16-CH), 6.97 (d, 1H, J=2.3 Hz, ArH), 7.01 (m, 1H, ArH),7.03 (s, 1H, NHC(CH₃)₃), 7.33 (d, 1H, J=8.6 Hz, ArH), 7.89 (s, 2H, NH₂).

Synthesis of 17-(N-Methylcarbamoyl)estra-1,3,5(10),16-tetraen-3-ylsulfamate (11f)

In similar manners to those described for the synthesis of sulfamate11a, compound 11f was prepared from compound 8. FAB-MS m/z 391 (M+H)⁺;¹H NMR (270 MHz, DMSO-d₆) δ 0.91 (s, 3H, CH₃), 2.62 (d, 3H, J=4.6 Hz,NHCH₃), 6.35 (s, 1H, 16-CH), 6.97 (s, 1H, ArH), 7.02 (d, 1H, J=8.6 Hz,ArH), 7.33 (d, 1H, J=8.6 Hz, ArH), 7.69 (d, 1H, J=4.6 Hz, NHCH₃), 7.88(s, 2H, NH₂).

Synthesis of 17-N-Ethylcarbamoyl)estra-1,3,5(10),16-tetraen-3-ylsulfamate (11g)

In similar manners to those described for the synthesis of sulfamate11a, compound 11g was prepared from compound 8. FAB-MS m/z 405 (M+H)⁺;¹H NMR (270 MHz, DMSO-d₆) δ 0.91 (s, 3H, CH₃), 1.03 (t, 3H, J=7.1 Hz,CH₂CH₃), 3.12 (m, 2H, CH₂CH₃), 6.36 (s, 1H, 16-CH), 6.97 (d, 1H, J=2.3Hz, ArH), 7.01 (dd, 1H, J=2.3, 8.6 Hz, ArH), 7.33 (d, 1H, J=8.6 Hz,ArH), 7.75 (t, 1H, J=5.7 Hz, NHCH₂), 7.89 (s, 2H, NH₂).

Synthesis of 17-(N-Isopropylcarbamoyl)estra-1,3,5(10),16-tetraen-3-ylsulfamate (11h)

In similar manners to those described for the synthesis of sulfamate11a, compound 11h was prepared from compound 8. FAB-MS m/z 419 (M+H)⁺;¹H NMR (270 MHz, DMSO-d₆) δ 0.91 (s, 3H, CH₃), 1.072 (d, 3H, J=5.6 Hz,CH(CH₃)₂), 1.074(d, 3H, J=6.6 Hz, CH(CH₃)₂), 3.93 (m, 1H, CH(CH₃)₂),6.36 (s, 1H, 16-CH), 6.97 (s, 1H, ArH), 7.01 (d, 1H, J=8.6 Hz, ArH),7.33 (d, 1H, J=8.6 Hz, ArH), 7.52 (m, 1H, NHCH), 7.88 (s, 2H, NH₂).

Synthesis of 17-(N-sec-Butylcarbamoyl)estra-1,3,5(10),16-tetraen-3-ylsulfamate (11i)

In similar manners to those described for the synthesis of sulfamate11a, compound 11i was prepared from compound 8. FAB-MS m/z 433 (M+MH)⁺;¹H NMR (270 MHz, DMSO-d₆) δ 0.82 (t, 3H, J=7.5 Hz, CH₂CH₃), 0.92 (s, 3H,CH₃), 1.04 (d, 3H, J=6.6 Hz, CHCH₃), 3.75 (m, 1H, CHCH₃), 6.34 (s, 1H,16-CH), 6.98 (s, 1H, ArH), 7.02 (d, 1H, J=8.6 Hz, ArH), 7.33 (d, 1H,J=8.6 Hz, ArH), 7.44 (m, 1H, NHCH), 7.88 (s, 2H, NH₂).

Synthesis of 17- N-tert-Amylcarbamoyl)estra-1,3,5(10),16-tetraen-3-ylsulfamate (11j)

In similar manners to those described for the synthesis of sulfamate11a, compound 11j was prepared from compound 8. FAB-MS m/z 447 (M+H)⁺;¹H NMR (270 MHz, DMSO-d₆) δ 0.76 (t, 3H, J=7.4 Hz, CH₂CH₃), 0.92 (s, 3H,CH₃), 1.22 (s, 3H, C(CH₃)₂), 1.23 (s, 3H, C(CH₃)₂), 6.29 (s, 1H, 16-CH),6.85 (m, 1H, NH), 6.98(s, 1H, ArH), 7.02 (d, 1H, J=8.6 Hz, ArH), 7.33(d, 1H, J=8.6 Hz, ArH), 7.87(s, 2H, NH₂).

Synthesis of 17-(N-Isoamylcarbamoyl)estra-1,3,5(10),16-tetraen-3-ylsulfamate (11k)

In similar manners to those described for the synthesis of sulfamate11a, compound 11k was prepared from compound 8. FAB-MS m/z 447 (M+H)⁺;¹H NMR (270 MHz, DMSO-d₆) δ 0.88 (d, 6H, J=6.6 Hz, CH(CH₃)₂), 0.91 (s,3H, CH₃), 3.10(m, 2H, NHCH₂), 6.33 (s, 1H, 16-CH), 6.97 (s, 1H, ArH),7.01 (d, 1H, J=8.6 Hz, ArH), 7.33 (d, 1H, J=8.6 Hz, ArH), 7.69 (t, 1H,J=5.6 Hz, NHCH₂), 7.88(s, 2H, NH₂).

Synthesis of 17-(N,N-Diethylcarbamoyl)estra-1,3,5(10),16-tetraen-3-ylsulfamate (11l)

In similar manners to those described for the synthesis of sulfamate11a, compound 11l was prepared from compound 8. TOF-MS m/z 433 (M+H)⁺;¹H NMR (270 MHz, DMSO-d₆) δ 0.99 (s, 3H, CH₃), 1.05 (t, 6H, J=6.9 Hz,CH₂CH₃), 3.20-3.48 (m, 4H, CH₂CH₃), 5.76-5.82 (m, 1H, 16-CH), 6.96 (d,1H, J=2.3 Hz, ArH), 7.01 (dd, 1H, J=2.3, 8.6 Hz, ArH), 7.31 (d, 1H,J=8.6 Hz, ArH), 7.84 (brs, 2H, NH₂).

Synthesis of17-(N-methyl-N-Propylcarbamoyl)estra-1,3,5(10),16-tetraen-3-yl sulfamate(11m)

In similar manners to those described for the synthesis of sulfamate11a, compound 11m was prepared from compound 8. TOF-MS m/z 433 (M+H)⁺;¹H NMR (270 MHz, DMSO-d₆) δ 0.81 (t, 3H, J=7.6 Hz, CH₂CH₃), 0.99 (s, 3H,CH₃), 2.74-3.04 (m, 3H, NCH₃), 3.20-3.40 (m, 2H, NCH₂), 5.81 (brs, 1H,16-CH), 6.96 (d, 1H, J=2.3 Hz, ArH), 7.00 (dd, 1H, J=2.3, 8.6 Hz, ArH),7.31 (d, 1H, J=8.6 Hz, ArH), 7.84 (brs, 2H, NH₂).

Synthesis of 17-(N-Ethoxycarbamoyl)estra-1,3,5(10),16-tetraen-3-ylsulfamate (11n)

According to the four steps shown in FIG. 4, compound 11n was preparedfrom compound 8a which was prepared from estrone acetate in accordancewith a known method (for example, Tetrahedron Letters26, 1109-1112,1985). TOF-MS m/z 421 is (M+H)⁺; ¹H NMR (270 MHz, DMSO-d₆) δ 0.92 (s,3H, CH₃), 1.15 (t, 3H, J=6.9 Hz, CH₂CH₃), 3.82 (q, 2H, J=7.2 Hz,CH₂CH₃), 6.29 (s, 1H, 16-CH), 6.98 (s, 1H, ArH), 7.02 (d, 1H, J=8.6 Hz,ArH), 7.33 (d, 1H, J=8.6 Hz, ArH), 7.88 (brs, 2H, NH₂), 10.93 (s, 1H,CONH).

Synthesis of 17-Carbamoylestra-1,3,5(10),16-tetraen-3-yl sulfamate (11o)

Compound 11o was synthesized in a similar manner as 11a except thatTMS-I was used in place of BBr₃. Compound 11o was prepared from compound12. EI-MS m/z 376 M⁺; ¹H NMR (300 MHz, DMSO-d₆) δ 0.88 (s, 3H, CH₃),6.44 (brs, 1H, 16-CH), 6.74(brs, 2H, CONH₂), 6.97 (s, 1H, ArH), 7.01 (d,1H, J=9 Hz, ArH), 7.32 (d, 1H, J=9 Hz, ArH), 7.89 (s, 2H, SO₂NH₂).

Synthesis of 17-(N,N-Dimethylcarbamoyl)estra-1,3,5(10),16-tetraen-3-ylsulfamate (11p)

Compound 11p was synthesized in a similar manner as 11a except thatTMS-I was used in place of BBr₃. Compound 11p was prepared from compound12. EI-MS m/z 404 M⁺; ¹H NMR (300 MHz, DMSO-d₆) δ 0.98 (s, 3H, CH₃),2.93 (brs, 6H, N(CH₃)₂), 5.86 (s, 1H, 16-CH), 6.97 (s, 1H, ArH), 7.02(d, 1H, J=9 Hz, ArH), 7.31 (d, 1H, J=9 Hz, ArH), 7.89 (s, 2H, SO₂NH₂).

Synthesis of17-(N-methyl-N-Methoxycarbamoyl)estra-1,3,5(10),16-tetraen-3-yl-sulfamate,(compound 11t)

Compound 11t was synthesized in a similar manner as 11a except thatTMS-I was used in place of BBr₃. Compound 11t was prepared from compound12. ¹H NMR (250 MHz, DMSO-d₆) δ 0.80 (s, 3H, CH₃), 2.98 (s, 3H, NCH₃),3.40 (s, 3H, OCH₃), 6.10(s, 1H, 16-CH), 6.95 (s, 1H, ArH), 7.15 (dd, 1H,J=8.4 Hz, ArH), 7.30 (d, 1H, J=8.4 Hz, ArH), 7.95 (s, 2H, NH₂).

Synthesis of17-(N,N-di-n-Propylcarbamoyl)estra-1,3,5(10),16-tetraen-3-yl sulfamate(compound 11q)

Compound 16q was obtained from compound 15 through carbonylinsertion-amidation; compound 11q was synthesized in a similar manner as11a except that TMS-I was used in place of BBr₃. EI-MS m/z 460 M⁺; ¹HNMR (250 MHz, DMSO-d₆) δ 0.90 (t, 6H, CH₂CH₃), 1.09 (s, 3H, CH₃),3.30-3.45 (m, 4H, NCH₂), 5.85 (s, 1H, 16-CH), 6.97 (s, 1H, ArH), 7.02(d, 1H, J=8.6 Hz, ArH), 7.36 (d, 1H, J=8.6 Hz, ArH), 7.95 (s, 2H,SO₂NH₂).

Synthesis of17-(N,N-diisopropylcarbamoyl)estra-1,3,5(10),16-tetraen-3-yl sulfamate(compound 11r)

Compound 16r was obtained from compound 15 through carbonylinsertion-amidation; compound 11r was synthesized in a similar manner as11a except that TMS-I was used in place of BBr₃. EI-MS m/z 460 M⁺; ¹HNMR (250 MHz, DMSO-d₆) δ 0.90 (s, 3H, CH₃), 1.30 (brs, 12H, CH(CH₃)₂),3.90 (m, 2H, CH(CH₃)₂), 5.45 (s, 1H, 16-CH) 6.75 (s, 1H, ArH), 6.85 (d,1H, J=8.6 Hz, ArH), 7.10 (d, 1H, J=8.6 Hz, ArH), 7.70 (s, 2H, SO₂NH₂).

Synthesis of17-(N-ethyl-N-Isopropylcarbamoyl)estra-1,3,5(10),16-tetraeu-3-ylsulfamate (compound 11s)

Compound 16s was obtained from compound 15 through carbonylinsertion-amidation; compound 11s was synthesized in a similar manner as11a except that TMS-I was used in place of BBr₃. EI-MS m/z 446 M⁺; ¹HNMR (250 MHz, DMSO-d₆) δ 1.20 (s, 3H, CH₃), 3.40 (m, 2H, CH₂CH₃), 4.45(brs, 1H, CH(CH₃)₂), 5.90 (s, 1H, 16-CH), 7.15 (s, 1H, ArH), 7.30 (d,1H, J=8.6 Hz, ArH), 7.50 (d, 1H, J=8.6 Hz, ArH), 8.05 (s, 2H, SO₂NH₂).

Example 3 Preparation of Class 3 Compounds

Reference numerals correspond with those shown in FIG. 7.

Synthesis of 3-Benzyloxy-17-(propylcarboxyl)estra-1,3,5(10),16-tetraene(Compound 18)

A mixture of triflate (compound 15) (1.1 g, 2.37 mmol), palladium (11)acetate (45 mg, 0.20 mmol), 1,3-Bis(diphenylphosphino)propane (dppp) (66mg, 0.16 mmol), and n-propanol (8 ml) in dimethylformamide (DMF, 10 ml)was heated at 60° C. with carbon monoxide bubbled through for 5.5 hours.The reaction mixture was then diluted with ethyl acetate and washed withwater. The organic layer was dried with sodium sulfate (Na₂SO₄),concentrated and the residue was purified by silica gel chromatographyand eluted with methylene chloride (CH₂Cl₂) yielding the pureα,β-unsaturated ester (compound 18) (875 mg, 89.1 %). m.p. 84-85° C.; ¹HNMR (300 MHz, CDCl₃) δ 0.97 (s, 3H, CH₃), 1.00 (t, 3H, J=7.5 Hz, CH₃),4.13 (t, 2H, 6.6 Hz, OCH₂), 5.05 (s, 2H, CH₂PH), 6.7-7.4 (m, 9H, 16-Hand ArH).

Synthesis of 3-Hydroxy-17-(propylcarboxyl)estra-1,3,5(10),16-tetraene(Compound 19)

Compound 18 (780 mg, 1.81 mmol) was dissolved in CH₂Cl₂ (16 ml) andtrimethylsilyl iodide (TMS-I, 0.9 ml, 6.28 mmol) was added in oneportion to the reaction mixture at room temperature and the solution wasstirred at room temperature for 15 min. Water was then added to thesolution and the mixture was stirred for 30 min. The mixture wasextracted with CH₂Cl₂ and the organic layer was separated and washedwith 10% sodium thiosulfate solution to neutralize the iodide and thesolution was dried with sodium sulfate (Na₂SO₄). The solution wasevaporated and the residue was purified by silica gel chromatography andeluted with methylene chloride (CH₂Cl₂):ethyl acetate (EtOAc) (4:1)yielding compound 19 (590 mg, 95.7%) m.p. 139.7-140.7° C.

Synthesis of 17-(n-Propylcarboxyl)estra-1,3,5(10),16-tetraene (Compound20a)

To a solution of compound 19 comprising 250 mg (0.73 mmol) of compound19 and 2,6di-tert-butyl-4-methylpyridine (DBMP) (0.6 g, 3.23 mmol) inCH₂Cl₂ (30 ml) was added sulfamoyl chloride (1 g, 8.6 mmol) portionwisewith stirring at 0° C. The solution was stirred for 3.5 hours at roomtemperature. The solution was washed with water until neutral, driedover Na₂SO₄, and evaporated under reduced pressure. The solid obtainedwas purified by chromatography (CH₂Cl₂/EtOAc, 4:1) to give 293 mg of 20(95%). m.p. 113.9-115.2° C.; ¹H NMR (300 MHz, DMSO-d₆) δ 0.89 (s, 3H,CH₃), 0.92 (t, 3H, J=7.5 Hz, CH₃), 4.04 (t, 2H, J=6.3 Hz, OCH₂), 6.78(brs, 1H, 16-CH), 6.98 (d, 1H, J=1.8 Hz, ArH), 7.02 (dd, 1H, J=1.8, 8.4Hz, ArH), 7.33 (d, 1H, J=8.4 Hz, ArH), 7.91 (s, 2H, NH₂). Analysiscalculated for C₂₂H₂₉NO₅S: C, 62.98; H, 6.97; N, 3.34. Found C, 62.86;H, 6.90; N, 3.32.

Synthesis of17-(Ethylcarboxyl)estra-1,3,5(10),16-tetraen-3-yl-sulfamate, (Compound20b)

Compound 20b was synthesized in a similar manner as compound 20a. ¹H NMR(250 MHz, DMSO-d₆) δ 0.95 (s, 3H, 18-CH₃), 1.20 (t, 3H, J=7.2 Hz, CH₃),4.10 (q, 2H, J=7.2 Hz, CH₂), 3.70 (s, 3H, CH₃), 6.70 (s, 1H, 16-CH),6.92 (s, 1H, ArH), 6.98 (dd, 1H, J=8.4 Hz, ArH), 7.30 (d, 1H, J=8.4 Hz,ArH), 7.82 (s, 2H, NH₂).

Synthesis of17-(Methylcarboxyl)estra-1,3,5(10),16-tetraen-3-yl-sulfamate, (Compound20c)

Compound 20c was synthesized in a similar manner as compound 20a. ¹H NMR(250 MHz, DMSO-d₆) δ 0.92 (s, 3H, CH₃), 3.70 (s, 3H, CH₃), 6.80 (s, 1H,16-CH), 6.98 (s, 1H, ArH), 7.0 (dd, 1H, J=1.8, 8.4Hz, ArH), 7.35 (d, 1H,J=8.4Hz, ArH), 7.90 (s, 2H, NH₂).

Example 4 Preparation of Class 2 Compounds

Reference numerals correspond with those shown in FIG. 11.

Synthesis of 17β-(N-Isobutylcarbamoyl)estra-1,3,5(10)-trien-3-ylsulfamate (Compound 29d)

In similar manners to those described for the synthesis of compound 7,compound 29d was prepared from compound 11d. TOF-MS m/z 435 (M+H)⁺; ¹HNMR (270 MHz, DMSO-d₆) δ 0.60 (s, 3H, CH₃), 0.84 (d, 6H, J=6.9 Hz,CH(CH₃)₂), 2.60-3.16 (m, 2H, NHCH₂), 6.95 (d, 1H, J=2.3 Hz, ArH), 7.00(dd, 1H, J=2.3, 8.6 Hz, ArH), 7.34 (d, 1H, J=8.6 Hz, ArH), 7.49 (brt,1H, J=5.6 Hz, NHCH₂), 7.84 (brs, 2H, NH₂).

Synthesis of 17β-(N-tert-Butylcarbamoyl)estra-1,3,5(10)-trien-3-ylsulfamate (Compound 29e)

In similar manners to those described for the synthesis of compound 7,compound 29e was prepared from compound 11e. FAB-MS m/z 435 (M+H)⁺; ¹HNMR (270 MHz, DMSO-d₆) δ 0.59 (s, 3H, CH₃), 1.26 (s, 9H, C(CH₃)₃), 6.89(s, 1H, NH),6.96 (d, 1H, J=2.3 Hz, ArH), 7.01 (dd, 1H, J=2.3, 8.6 Hz,ArH), 7.34 (d, 1H, J=8.6 Hz, ArH), 7.87 (s, 2H, NH₂).

Synthesis of 17β-(N-Methylcarbamoyl)estra-1,3,5(10)-trien-3-yl sulfamate(Compound 29f)

In similar manners to those described for the synthesis of compound 7,compound 29f was prepared from compound 11f. FAB-MS m/z 393 (M+H)⁺; ¹HNMR (270 MHz, DMSO-d₆) δ 0.60 (s, 3H, CH₃), 2.59 (d, 3H, J=3.6 Hz,NHCH₃), 6.96 (d, 1H, J=2.3 Hz, ArH), 7.01 (dd, 1H, J=2.3, 8.6 Hz, ArH),7.34 (d, 1H, J=8.6 Hz, ArH), 7.29 (m, 1H, NHCH₃), 7.87 (s, 2H, NH₂).

Synthesis of 17β-(N-Ethylcarbamoyl)estra-1,3,5(10)-trien-3-yl sulfamate(Compound 29g)

In similar manners to those described for the synthesis of compound 7,compound 29g was prepared from compound 11g. FAB-MS m/z 407 (M+H)⁺; ¹HNMR (270 MHz, DMSO-d₆) δ 0.60 (s, 3H, CH₃), 1.01 (t, 3H, J=7.3 Hz,CH₂CH₃), 3.02 (m, 1H, CH₂CH₃), 3.16 (m, 1H, CH₂CH₃), 6.96 (d, 1H, J=2.0Hz, ArH), 7.01 (d, 1H, J=8.6 Hz, ArH), 7.34 (d, 1H, J=8.6 Hz, ArH), 7.47(t, 1H, J=5.6 Hz, NHCH₂), 7.87 (s, 2H, NH₂).

Synthesis of 17β-(N-Isopropylcarbamnoyl)estra-1,3,5(10)-trien-3-ylsulfamate (Compound 29h)

In similar manners to those described for the synthesis of compound 7,compound 29h was prepared from compound 11h. TOF-MS m/z 421 (M+H)⁺; ¹HNMR (270 MHz, DMSO-d₆) δ 0.58 (s, 3H, CH₃), 1.00-1.10 (m, 6H, CH(CH₃)₂),3.38-3.50 (m, 1H, CH(CH₃)₂), 6.95 (d, 1H, J=2.3 Hz, ArH), 7.00 (dd, 1H,J=2.3, 8.6 Hz, ArH), 7.27 (d, 1H, J=7.9 Hz, NHCH), 7.33 (d, 1H, J=8.6Hz, ArH), 7.85 (brs, 2H, NH₂).

Synthesis of 17β-(N-methyl-N-Propylcarbamoyl)estra-1,3,5 (10)-trien-3-ylsulfamate (Compound 29m)

In similar manners to those described for the synthesis of compound 7,compound 29m was prepared from compound 11m. TOF-MS m/z 435 (M+H)⁺; ¹HNMR (270 MHz, DMSO-d₆) δ 0.60-0.68 (m, 3H, CH₃), 0.76-0.88 (m, 3H,CH₂CH₃), 2.70-3.70 (m, 5H, N(CH₃)CH₂), 6.95 (d, 1H, J=2.6 Hz, ArH), 7.00(dd, 1H, J=2.6, 8.6 Hz, ArH), 7.32 (d, 1H, J=8.6 Hz, ArH), 7.83 (brs,2H, NH₂).

Example 5 Preparation of Class 10 Compounds

Reference numerals correspond with those shown in FIG. 9.

Synthesis of 17-(N-n-Propycarbamoyl)-estra-1,3,5(10),16-tetraen-[3,2,e]-1′2′3′-oxathiazine-2,2′-dioxide (Compound 22)

To a solution of compound 21 (0.37 g, 1.0 mmol) in DMF (10 ml) was addedsodium hydride (0.115 g, 5 mmol) at (0° C. The reaction mixture wasstirred for 20 minutes and chlorosulfonamide (0.73 g, 5 mmol) was added.The solution was stirred for an additional 6 hrs. The reaction mixturewas poured into ice cold saturated NH₄Cl solution and then extractedwith EtOAc. The organic layer was separated, washed with water, brineand dried (Na₂SO₄). The solvent was evaporated under reduced pressureand the residue was purified by silica gel chromatography (CH₂Cl₂;EtOAc, 15:1). FAB-MS m/z 428 (M+H)⁺; NMR (250 MHz, DMSO-d₆) δ 0.65 (t,3H, CH₂CH₃), 0.70 (s, 3H, CH₃), 6.15 (s, 1H, 16-CH), 7.01 (s, 1H, ArH),7.55 (m, 1H, NH), 7.70 (s, 1H, ArH), 8.90 (s, 1H—CH═N—).

Synthesis of 17-(N-n-Propylcarbamoyl)-estra-1,3,5(10),16-tetraen-[3,2,e]-3′4′-dihydro-1′2′3′-oxathiazine-2,2′-dioxde (Compound 23)

To a solution of compound 22 (0.428 g, 1mmol) in methanol (4 ml) wasadded NaBH₄ (0.037 g, 0.96 mmol) at 0° C. The reaction was stirred at 0°C. for 1 hr and the mixture was poured into saturated NH₄Cl andextracted with EtOAc. The organic layer was separated, washed withwater, brine and died (Na₂SO₄) and the solvent was evaporated underreduced pressure. The residue was purified by silica gel chromatography(EtOAc: Pet ether, 1:1). FAB-MS m/z 430 (M+H)⁺; ¹H NMR (250 MHz,DMSO-d₆) δ 0.65 (t, 3H, CH₂CH₃), 0.75 (s, 3H, CH₃), 4.30 (s, 2H,CH₂NHSO₂), 6.20 (s, 1H, 16-CH), 6.55 (s, 1H, ArH), 7.05 (s, 1H, ArH),7.60 (m, 1H, amide NH), 8.25 (m, 1H, CH₂NHSO₂).

Example 6 Preparation of Class 6 Compounds

Reference numerals correspond with those shown in FIG. 12.

Synthesis of 17β-(Butyrylamino)estra-1,3,5(10)-trien-3-yl sulfamate(Compound 30)

Compound 30 was prepared from compound 24 in three steps (amidation,deprotection and sulfamoylation) according to the method taught by Li,et al. Steroids, 63:425-32 (1998). FAB-MS m/z 421 (M+H)⁺; ¹H NMR (270MHz, DMSO-d₆) δ 0.65 (s, 3H, CH₃), 0.86 (t, 3H, J=7.6 Hz, CH₂CH₃), 2.07(m, 2H, COCH₂), 3.81 (m, 1H, 17-CH), 6.96 (d, 1H, J=2.3 Hz, ArH), 7.00(d, 1H, J=8.6 Hz, ArH), 7.32 (d, 1H, J=8.6 Hz, ArH), 7.48 (d, 1H, J=8.9Hz, NHCO), 7.87 (s, 2H, NH₂).

Example 7

The compounds prepared according to the methods of the above Exampleswere tested for biological activity in inhibiting sulfatase activityusing an in vitro conversion assay procedure. The specific compoundstested are shown in Table 2, below. As will be appreciated by thoseskilled in the art, this assay is based on inhibition of the conversionof ³H-estrone sulfate to ³H-estrone by the enzyme estrone sulfatase. Thefinal volume of the enzyme assay was 0.15 ml. [6.7-³H] estrone sulfate(final concentration 3.3 nmole/L; 300,000 dpm/tube); an inhibitor atvarious concentrations in DMSO and recombinant human estrone sulfatase(33 ng/tube) in phosphate buffered saline, containing 0.25 M sucrose and0.04 M nicotinamide, pH 7, were added to a 1.5 ml microtube. Recombinanthuman estrone sulfatase was partially purified from Chinese HamsterOvary (CHO) cells in which human estrone sulfatase cDNA was transfected.The assay began by the addition of the substrate estrone sulfate. After60 minutes of incubation at 37° C., 0.5 ml of toluene were added toquench the assay. Control samples with no inhibitor were incubatedsimultaneously. Blank samples were obtained by incubating withoutestrone sulfatase. The quenched samples were vortexed for 30 seconds andcentrifuged (9,000 rpm for 5 minutes). Two hundred and fifty microlitersof toluene were obtained from each quenched sample to determine theamount of product formation. Product formation for samples containing aninhibitor was compared to that of control samples (without inhibitors)run simultaneously. This was reported as percent inhibition of controlsample which equals

100× Product formation for sample containing inhibitor Product formationfor sample with no inhibitor (control)

Each compound was tested over a range of concentrations (0.025 to 2.5nM) for determination of dose-responsiveness and for calculation of IC₅₀values. Each concentration was tested in duplicate in three separateexperiments. Both compounds showed dose-dependent inhibition of estronesulfatase activity as determined by the in vitro conversion assay. TheIC₅₀ values, representing the concentration that resulted in 50%inhibition of estrone sulfatase activity in this assay, are shown inTable 2. The IC₅₀ values were determined by performing linear regressionanalysis of percent of control versus concentration (log 10); theresulting equation was then used to determine the concentration thatresulted in 50% inhibition.

TABLE 2

R₃ IC₅₀(nmol/L)

12

14

5

12

12

12

13

50

10

12

3.2

3.5

3.6

2.5

4.8

Example 8

Compounds 7, 29d, 29h and 29m were tested in the manner described forExample 6. Results are presented in Table 3.

TABLE 3

R₃ IC₅₀(nmol/L)

19

6.5

17

5.5

Whereas particular embodiments of this invention have been describedabove for purposes of illustration, it will be evident to those skilledin the art that numerous variations of the details of the presentinvention may be made withoutdeparting from the invention as defined inthe appended claims.

What is claimed is:
 1. A method of treating a patient for an estrogendependent illness comprising administering to said patient an effectiveamount of a compound having the formula: wherein R₁ and R₂ areindependently selected from hydrogen and a lower alkyl group having oneto six carbons; R₃ is:  R₄ and R₅ are independently selected fromhydrogen, straight or branched chain alkyl groups having one to fourteencarbons, and straight or branched chain alkoxy groups having one to sixcarbons and wherein X and Y are both carbons and the bond between X andY is either single or double.
 2. The method of claim 1, wherein R₁ andR₂ are both hydrogen and the bond between X and Y is double.
 3. Themethod of claim 2, wherein R₄ is H and R₅ is selected from the groupconsisting of hydrogen and methyl, ethyl, propyl, isopropyl, isobutyl,tert-butyl, sec-butyl, isopentyl, hexyl and ethoxy groups.
 4. The methodof claim 2, wherein R₄ is an ethyl group and R₅ is selected fromthegroup consisting of ethyl and isopropyl groups.
 5. The method ofclaim 2, wherein R₄ is methyl and R₅ is methyl or propyl.
 6. The methodof claim 2, wherein R₄ and R₅ are both methyl groups, both propyl groupsor both isopropyl groups.
 7. The method of claim 1, wherein R₁ and R₂are both hydrogen and the bon between X and Y is single.
 8. The methodof claim 7, wherein R₄ is H and R₅ is selected from the group consistingof methyl, ethyl, propyl, isopropyl, isobutyl and tert-butyl groups. 9.The method of claim 7, wherein R₄ is a methyl group and R₅ is a propylgroup.
 10. The method of claim 1, wherein said compound is containedwithin a suitable pharmaceutical carrier.
 11. The method of claim 1,wherein said administration is oral.
 12. The method of claim 1, whereinsaid effective amount is either a therapeutially effective amount, or aprophylactically effective amount.
 13. A compound having the formula

wherein R₁ and R₂ are independently selected from hydrogen and a loweralkyl group having one to six carbons; and

R₈ is hydrogen, and R₉ is a straight or branched alkanoyl group havingone to eight carbons.
 14. A compound of claim 13, wherein R₉ isCOCH₂CH₂CH₃.
 15. A method of treating a patient for an estrogendependent illness comprising administering to said patient an effectiveamount of a compound having the formula:

wherein R₁ and R₂ are independently selected from hydrogen and a loweralkyl group having one to six carbons; and R₃ is

R₈ is hydrogen, and R₉ is a straight or branched alkanoyl groups havingone to right carbons.
 16. The method of claim 15, wherein said compoundis contained within a suitable pharmaceutical carrier.
 17. The method ofclaim 15, wherein said administration is oral.
 18. The method of claim15, wherein said effective amount is either a therapeutically effectiveamount, or a prophylactically effective amount.
 19. A method of claim15, wherein R₉ is COCH₂CH₂CH₃.